For label-free experiments, Jurkat E6-1 cells (ATCC) were grown in Roswell Park Memorial Institute 1640 (RPMI) media (Gibco) supplemented with 10% fetal bovine serum (FBS) (Gibco), penicillin, and streptomycin (Gibco). For SILAC experiments human embryonic kidney 293T (HEK293) cells transfected with … See more Three biological replicates of SILAC-labeled HEK293T cells were labeled with biotin-phenol (BP) (Iris Biotech GmbH) at >80% confluency using previously described methods4. … See more Jurkat cells were lysed with 8 M urea, 50 mM Tris-HCl pH 8.0, 150 mM NaCl, 1 mM EDTA (Sigma), 2 μg/μl Aprotinin (Sigma-Aldrich), 10 μg/μl Leupeptin (Roche), and 1 mM phenylmethylsulfonyl fluoride (PMSF) (Sigma) at 4 … See more A polyclonal anti-biotin (ImmuneChem Pharmaceuticals Inc., cat no. ICP0615) was used for biotin peptide enrichment. Peptide spike-in samples were composed of varying amounts of biotinylated peptides spiked in to … See more Jurkat peptides were labeled with EZ link NHS-Biotin (Thermo Fisher Scientific). The biotin reagent was brought to RT. For labeling, 1 mg of peptide from Jurkat cell lysate was … See more
Optimized APEX2 peroxidase-mediated proximity labeling in fast
WebAPEX2-MYC quantitative mass spectrometry: U2OS expressing an APEX2-MYC fusion protein were incubated with medium containing 1mM Biotin-Phenol (Iris Biotech) for 2 h at 37°C. H2O2 was added to a final concentration of 1mM and incubated for 2 min. Biotinylation was stopped by washing three times with quenching solution (10 mM … WebDec 18, 2024 · Alk-Ph is more water soluble and cell wall permeable than biotin-phenol substrate, allowing more efficient profiling of the subcellular proteome in microorganisms. We describe the protocol for Alk-Ph probe synthesis, APEX2 expression, and protein/RNA labeling in yeast and the workflow for quantitative proteomic experiments and data analysis. green lake festival of music 2022
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WebIris Biotech GmbH Biotin-AEEA-phenol Revision date: 12.01.2024 Product code: LS-3490 Page 6 of 7 No data available Specific effects in experiment on an animal No data available Practical experience 11.2. Information on other hazards Further information RTECS: No data available Caution! To the best of our knowledge the toxicological properties ... WebOct 16, 2024 · Biotin-phenol labeling. Three biological replicates of SILAC-labeled HEK293T cells were labeled with biotin-phenol (BP) (Iris Biotech GmbH) at >80% confluency using previously described methods 4 ... WebSynonym(s): (3aS,4S,6aR)-Hexahydro-N-[2-(4-hydroxyphenyl)ethyl]-2-oxo-1H-thieno[3,4-d]imidazole-4-pentanamide, BP, Biotin-Phenol. Empirical Formula (Hill Notation): C 18 … green lake fishing seattle